• Title/Summary/Keyword: dsDNA

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STING Negatively Regulates Double-Stranded DNA-Activated JAK1-STAT1 Signaling via SHP-1/2 in B Cells

  • Dong, Guanjun;You, Ming;Ding, Liang;Fan, Hongye;Liu, Fei;Ren, Deshan;Hou, Yayi
    • Molecules and Cells
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    • v.38 no.5
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    • pp.441-451
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    • 2015
  • Recognition of cytosolic DNA initiates a series of innate immune responses by inducing IFN-I production and subsequent triggering JAK1-STAT1 signaling which plays critical roles in the pathogenesis of infection, inflammation and autoimmune diseases through promoting B cell activation and antibody responses. The stimulator of interferon genes protein (STING) has been demonstrated to be a critical hub of type I IFN induction in cytosolic DNA-sensing pathways. However, it still remains unknown whether cytosolic DNA can directly activate the JAK1-STAT1 signaling or not. And the role of STING is also unclear in this response. In the present study, we found that dsDNA directly triggered the JAK1-STAT1 signaling by inducing phosphorylation of the Lyn kinase. Moreover, this response is not dependent on type I IFN receptors. Interestingly, STING could inhibit dsDNA-triggered activation of JAK1-STAT1 signaling by inducing SHP-1 and SHP-2 phosphorylation. In addition, compared with normal B cells, the expression of STING was significantly lower and the phosphorylation level of JAK1 was significantly higher in B cells from MRL/lpr lupus-prone mice, highlighting the close association between STING low-expression and JAK1-STAT1 signaling activation in B cells in autoimmune diseases. Our data provide a molecular insight into the novel role of STING in dsDNA-mediated inflammatory disorders.

Isolation and Identification of a Biphenyl-degrading Bacterium, Pseudomonas sp. DS-94 (Biphenyl 분해 미생물 Pseudomonas sp. DS-94의 분리 및 동정)

  • Lee, Dae-Sung;Jeong, Seong-Yun
    • Journal of Environmental Science International
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    • v.19 no.11
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    • pp.1391-1396
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    • 2010
  • Three biphenyl-degrading microorganisms were isolated from polluted soil samples in Sasang-gu, Busan. Among them, isolate DS-94 showing the strong degrading activity was selected. The morphological, physiological, and biochemical characteristics of DS-94 were investigated by API 20NE and other tests. This bacterium was identified as the genus Pseudomonas by 16S rDNA sequencing and designated as Pseudomonas sp. DS-94. The optimum temperature and pH for the growth of Pseudomonas sp. DS-94 were $25^{\circ}C$ and pH 7.0, respectively. This isolate could utilize biphenyl as sole source of carbon and energy. Biphenyl-degrading efficiency of this isolate was measured by HPLC analysis. As a result of biological biphenyl-degradation at high biphenyl concentration (500 mg/L), biphenyl-removal efficiency by this isolate was 73.5% for 7 days.

The Genera Babuvirus and Badnavirus in Asia

  • Natsuaki, Keiko T.;Furuya, Noriko
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.227-232
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    • 2007
  • In the plant virus world, there are six genera of plant viruses with dsDNA genomes and six genera with ssDNA (Fauquet et al., 2005). The dsDNA viruses are comprised of 4 genera in the Caulimoviridae, the genus Badnavirus and the genus Tungrovirus. The ssDNA viruses are comprised of four genera in Geminiviridae, and the two genera Nanovirus and Babuvirus in the Nanoviridae. The genera Babuvirus and Badnavirus are not well studied in Asia. However, we recognized the significance of two species, Banana bunchy top virus (BBTV) in the genus Babuvirus and Banana streak virus (BSV) in the genus Badnavirus, during the survey of banana viruses in Asia. Their main characters will be introduced in this mini-review.

Quantitative Approaches to Assess Key Carcinogenic Events of Genotoxic Carcinogens

  • Fukushima, Shoji;Gi, Min;Fujioka, Masaki;Kakehashi, Anna;Wanibuchi, Hideki;Matsumoto, Michiharu
    • Toxicological Research
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    • v.34 no.4
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    • pp.291-296
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    • 2018
  • Chemical carcinogenesis is a multistep process. Genotoxic carcinogens, which are DNA-reactive, induce DNA adduct formation and genetic alterations in target cells, thereby generating mutated cells (initiation). Subsequently, preneoplastic lesions appear through clonal proliferation of the mutated cells and transform into tumors (promotion and progression). Many factors may influence these processes in a dose-dependent manner. Therefore, quantitative analysis plays an important role in studies on the carcinogenic threshold of genotoxic carcinogens. Herein, we present data on the relationship between key carcinogenic events and their deriving point of departure (PoD). Their PoDs were also compared to those of the carcinogenesis pathway. In an experiment, the liver of rats exposed to 2-amino-3,8-dimethylimidazo-(4,5-f)quinoxaline (MeIQx) was examined to determine the formation of MeIQx-DNA adducts, generation of mutations at LacI transgene, and induction of preneoplastic glutathione S-transferase placental form (GST-P)-positive foci and tumors (benign and malignant). The PoDs of the above key events in the carcinogenicity of MeIQx were increased as the carcinogenesis advanced; however, these PoDs were lower than those of tumor induction. Thus, the order of key events during tumor induction in the liver was as follows: formation of DNA adducts ${\ll}$ Mutations ${\ll}$ GST-positive foci (preneoplasia) ${\ll}$ Tumor (adenoma and carcinoma). We also obtained similar data on the genotoxic and carcinogenic PoDs of other hepatocarcinogens, such as 2-amino-3,8-dimethylimidazo(4,5-f)quinoline. These results contribute to elucidating the existence of a genotoxic and carcinogenic threshold.

Ac/Ds-mediated gene tagging system in rice

  • Eun, Moo-Young;Yun, Doh-Won;Nam, Min-Hee;Yi, Gi-Hwan;Han, Chang-Deok;Kim, Doh-Hoon;Park, Woong-June;Kim, Cheol-Soo;Park, Soon-Ki
    • Proceedings of the Korean Society of Plant Biotechnology Conference
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    • 2005.11a
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    • pp.95-105
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    • 2005
  • Transposon-mediated insertional mutagenesis provides one of the most powerful tools for functional studies of genes in higher plants. This project has been performed to develop a large population of insertional mutations, and to construct databases of molecular information on Ds insertion sites in rice. Ultimate goals are to supply genetic materials and information to analyze gene function and to identify and utilize agronomically important genes for breeding purpose. Two strategies have been employed to generate the large scale of transposon population in a Japonica type rice, Dongjin Byeo; 1) genetic crosses between Ac and Ds lines and 2) plant regeneration from seeds carrying Ac and Ds. Our study showed that over 70% of regenerated plants generally carried independent Ds elements and high activity of transposition was detected only during regeneration period. Ds-flanking DNA amplified from leaf tissues of F2 and T1 (or T2) plants have been amplified via TAIL-PCR and directly sequenced. So far, over 65,000 Ds lines have been generated and over 9,500 Ds loci have been mapped on chromosomes by sequence analysis. Database of molecular information on Ds insertion sites has been constructed, and has been opened to the public and will be updated soon at http://www.niab.go.kr. Detailed functional analysis of more than 30 rice mutants has been performed. Several Ds-tagged rice genes that have been selected for functional analysis will be briefly introduced. We expect that a great deal of information and genetic resources of Ds lines would be obtained during the course of this project, which will be shared with domestic and international rice researchers. In addition to the Japonica rice, we have established the tagging system in an rice line of indica genetic background, MGRI079. MGRI079 (Indica/Japonica) was transformed with Agrobacteria carrying Ac and Ds T-DNA vectors. Among transgenic lines, we successfully identified single-copy Ds and Ac lines in MGR1079. These lines were served as ‘starter lines’ to mutagenize Indica genetic background. To achieve rapid, large scale generation of Ds transposant lines, MGR1079 transformants carrying homozygous Ac were crossed with ones with homozygous Ds, and $F_2$seeds were used for plant regeneration. In this year, over 2,000 regeneration plants were grown in the field. We are able to evaluate the tagging efficiency in the Indica genetic background in the fall.

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Newly developed MSAP analysis reveals the different polymorphism patterns in transgenic tobacco plants with the dsRNA MET1 gene

  • Oh, Yun Jung;Chung, Hee;Yu, Jae Gyeong;Park, Young Doo
    • Plant Biotechnology Reports
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    • v.3 no.2
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    • pp.139-145
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    • 2009
  • DNA methylation is known to play an important role in the regulation of gene expression in eukaryotes. In this study, we isolated NtMET1 from Nicotiana tabacum cv. Havana (SR1) and obtain transgenic plants that reduced MET1 expression level with the double-strand RNA (dsRNA) MET1 gene. Transgenic tobacco plants showed dwarf and abnormal flower development when compared with the wild type. Using methylation-sensitive amplified polymorphism (MSAP) analysis, the patterns of cytosine methylation in transformed plants and the wild type were compared. MseI/HpaII selection primers showed an interesting polymorphism, and 153 DNA bands of interest were detected. Among these, 30 selective fragments were sequenced and analyzed with a BLAST search by successful MSAP modifications. The homology search showed that the transposons and tandem repeated sequences were related to the phenotypes. These results suggested that the decreased degree of methylation by dsRNA strategy caused abnormal growth and development in N. tabacum.

Detection of Innate and Artificial Mitochondrial DNA Heteroplasmy by Massively Parallel Sequencing: Considerations for Analysis

  • Kim, Moon-Young;Cho, Sohee;Lee, Ji Hyun;Seo, Hee Jin;Lee, Soong Deok
    • Journal of Korean Medical Science
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    • v.33 no.52
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    • pp.337.1-337.14
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    • 2018
  • Background: Mitochondrial heteroplasmy, the co-existence of different mitochondrial polymorphisms within an individual, has various forensic and clinical implications. But there is still no guideline on the application of massively parallel sequencing (MPS) in heteroplasmy detection. We present here some critical issues that should be considered in heteroplasmy studies using MPS. Methods: Among five samples with known innate heteroplasmies, two pairs of mixture were generated for artificial heteroplasmies with target minor allele frequencies (MAFs) ranging from 50% to 1%. Each sample was amplified by two-amplicon method and sequenced by Ion Torrent system. The outcomes of two different analysis tools, Torrent Suite Variant Caller (TVC) and mtDNA-Server (mDS), were compared. Results: All the innate heteroplasmies were detected correctly by both analysis tools. Average MAFs of artificial heteroplasmies correlated well to the target values. The detection rates were almost 90% for high-level heteroplasmies, but decreased for low-level heteroplasmies. TVC generally showed lower detection rates than mDS, which seems to be due to their own computation algorithms which drop out some reference-dominant heteroplasmies. Meanwhile, mDS reported several unintended low-level heteroplasmies which were suggested as nuclear mitochondrial DNA sequences. The average coverage depth of each sample placed on the same chip showed considerable variation. The increase of coverage depth had no effect on the detection rates. Conclusion: In addition to the general accuracy of the MPS application on detecting heteroplasmy, our study indicates that the understanding of the nature of mitochondrial DNA and analysis algorithm would be crucial for appropriate interpretation of MPS results.

Complete Genome Sequence of Bifidobacterium bifidum DS0908, Isolated from Human Fecal Sample

  • Haneol Yang;Yong-Sik Kim;Doo-Sang Park
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.566-568
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    • 2023
  • In this report, we present the whole-genome sequence of Bifidobacterium bifidum DS0908 isolated from the human fecal sample. The genome composed of a single circular chromosome is 2,223,317 bp long and the DNA G+C content is 62.65%. No virulence genes were detected in the genomic sequences of B. bifidum DS0908.

Cohesion Establishment Factors Stimulate Endonuclease Activity of hFen1 Independently and Cooperatively

  • Kim, Do-Hyung;Kim, Jeong-Hoon;Park, Byoung Chul;Cho, Sayeon;Park, Sung Goo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1768-1771
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    • 2015
  • Human Fen1 protein (hFen1) plays an important role in Okazaki fragment processing by cleaving the flap structure at the junction between single-stranded (ss) DNA and doublestranded (ds) DNA, an intermediate formed during Okazaki fragment processing, resulting in ligatable nicked dsDNA. It was reported that hChlR1, a member of the cohesion establishment factor family, stimulates hFen1 nuclease activity regardless of its ATPase activity. In this study, we found that cohesion establishment factors cooperatively stimulate endonuclease activity of hFen1 in in vivo mimic condition, including replication protein-A-coated DNA and high salt. Our findings are helpful to explain how a DNA replication machinery larger than the cohesion complex goes through the cohesin ring structure on DNA during S phase in the cell cycle.